4. Hrvatski kongres kliniËke citologije 4th Croatian Congress ... - Penta

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4. Hrvatski kongres kliniËke citologije / 1. Hrvatski simpozij analitiËke citologije / 2. Hrvatski simpozij citotehnologije IMMUNODETECTION OF ANTIGENES IN CYTOLOGIC SAMPLES Rešetar R University Hospital Center Rijeka, Croatia Objective: Presentation of the procedures of antigene detection in cytologic smears depending on their cell location. Methods: Basic rules of immunodetection methods have been used in the research. Cell antigens can be divided into three basic groups: membrane, cytoplasmic and nuclear antigens. Since membrane and cytoplasmic antigens are more accessible to the reaction with specific antibody due their cell location, there were no difficulties in their detection. However, nuclear antigens, especially Ki-67 placed deep in the cell nucleus, can not be detected just by using classic procedures of immunocytochemistry. Due to its specific location in the cell and in order to prove nuclear antigens, an additional special technique called ‘antigen demasking’ is needed. This technique is performed with the help of special fluid for ‘antigen demasking’ and thermic treatment in the water bath. In immunocytochemical methods there are three key steps important for the validity and positive result of reactions: 1. preparation fixation time; 2. precisely performed ‘antigen demasking’; 3. incubation time for primary monoclonal antibody. In immunocytochemical methods we have combined the following procedures:1.fixation time: 2-10 min, 2.’antigen demasking’ time: 5-20 min, 3. fluid for ‘antigen demasking’: citrate buffer (ph=6) or Tris/EDTA (ph=9),4. incubation time for primary monoclonal antibody: 30-90 min. For all types of antigens we used sylanized glass impregnated by special adhesion matters. Results: Membrane and cytoplasmic antigens are detected by classic procedures of immunocytochemistry with 10 minute fixation and 30-60 minute incubation with antibodies. The following procedure showed the best results for the detection of nuclear antigens: 10 minute fixation in cold acetone, 5 minute incubation in water bath on 95 °C in citrate buffer (ph=6) and 90 minute incubation with primary antibody. Conclusion: For proving specific nuclear antigens all four steps are needed as well as the procedure of ‘antigene demasking’ which includes thermic treatment in water bath with obligatory use of fluid for ‘antigene demasking’. Membrane and cytoplasmic antigens require only the combination of fixation and adequate incubation time. nale.kralj@gmail.com 178 Citotehnologija - Usmena predavanja
Cytotechnology - Oral Presentations INFLUENCE OF INADEQUATE MATERIALS ON CYTOLOGICAL ANALISIS AND PROVIDING DIAGNOSIS Rubić K, Milanković Lj, Horvat B, Ropar S General Hospital Karlovac, Karlovac, Croatia Identification of disease and providing diagnosis are often hard to establish because of inadequate material. More and more is the case that exfoliative materials which are often inadequate to examine are brought to a cytological laboratory. In some cases materials are not brought on time, so their analysis is not possible. Some materials are not usable medium for cells and longer air-contact and holding on warm area cause decay of cells and replication of bacteria, which all influence cell-analysis. Such materials are hard to analyse, most of them even impossible. All specimens whol centrifugate in cytocentrifuge-cytospin and painted with May-Grunwald stain. Specimens are repeated many times.Repeating the specimens more slowly, malign diagnosis have been noticed with some of them. In order to get better quality, faster and exacter diagnosis, better staff-education and team-communication are among persons who work on taking and distribuing materials in purpose of removal of causes are needed. That would minimize mistakes, diagnosis would be faster and exacter and the number of controls would be smaller. katica.rubic@ka.t-com.hr 179 4 th Croatian Congress of Clinical Cytology / 1 st Croatian Symposium of Analytical Cytology / 2 nd Croatian Symposium of Cytotechnology
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Final programme and abstract book
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UTORAK, 13.10.2009. SRIJEDA, 14.10.
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4. Hrvatski kongres kliniËke citologije 4th Croatian Congress ... - Penta

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